WARNING: This product is for research use only, not for human or veterinary use.
MedKoo CAT#: 205892
CAS#: 1380288-87-8
Description: Pinometostat, also known as EPZ-5676, is a small molecule inhibitor of histone methyltransferase with potential antineoplastic activity. Upon intravenous administration, EPZ-5676 specifically blocks the activity of the histone lysine-methyltransferase DOT1L, thereby inhibiting the methylation of nucleosomal histone H3 on lysine 79 (H3K79) that is bound to the mixed lineage leukemia (MLL) fusion protein which targets genes and blocks the expression of leukemogenic genes.
MedKoo Cat#: 205892
Name: Pinometostat
CAS#: 1380288-87-8
Chemical Formula: C30H42N8O3
Exact Mass: 562.33799
Molecular Weight: 562.71
Elemental Analysis: C, 64.03; H, 7.52; N, 19.91; O, 8.53
Synonym: EPZ5676; EPZ-5676; EPZ 5676; Pinometostat.
IUPAC/Chemical Name: (2R,3R,4S,5R)-2-(6-amino-9H-purin-9-yl)-5-((((1r,3S)-3-(2-(5-(tert-butyl)-1H-benzo[d]imidazol-2-yl)ethyl)cyclobutyl)(isopropyl)amino)methyl)tetrahydrofuran-3,4-diol
InChi Key: LXFOLMYKSYSZQS-LURJZOHASA-N
InChi Code: InChI=1S/C30H42N8O3/c1-16(2)37(13-22-25(39)26(40)29(41-22)38-15-34-24-27(31)32-14-33-28(24)38)19-10-17(11-19)6-9-23-35-20-8-7-18(30(3,4)5)12-21(20)36-23/h7-8,12,14-17,19,22,25-26,29,39-40H,6,9-11,13H2,1-5H3,(H,35,36)(H2,31,32,33)/t17-,19+,22-,25-,26-,29-/m1/s1
SMILES Code: O[C@H]1[C@H](N2C=NC3=C(N)N=CN=C23)O[C@H](CN([C@H]4C[C@@H](CCC5=NC6=CC(C(C)(C)C)=CC=C6N5)C4)C(C)C)[C@H]1O
Appearance: Solid powder
Purity: >99% (or refer to the Certificate of Analysis)
Shipping Condition: Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition: Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility: Soluble in DMSO, not in water
Shelf Life: >2 years if stored properly
Drug Formulation: Stock solutions (50 or 10 mM) were prepared in dimethylsulfoxide (DMSO) and stored at −20°C.
Stock Solution Storage: 0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code: 2934.99.9001
| Biological target: | Pinometostat (EPZ-5676) is a DOT1L histone methyltransferase inhibitor with a Ki of 80 pM. |
| In vitro activity: | EPZ5676, a DOT1L inhibitor approved for use in clinical trials, is less toxic and more effective than EPZ004777. Here, this study found that EPZ5676 was more efficient than EPZ004777 in down-regulating HOXA9, PBX3 and H3K79m2 expression in human NPMc+ cells (Figure S5A), and in promoting cell apoptosis (Figure S5B). EPZ5676 gradually reduced HOXA9 and PBX3 expression in OCI-AML3 cells as the treatment time increased. The levels of cleaved caspase 3 and PARP were also increased by EPZ5676 treatment in a time-dependent manner (Figure 5A). These changes were not observed in OCI-AML2 cells (Figure 5A). The two cell lines were treated with EPZ5676 for 3 days or 7 days and then were stained with Annexin V/PI for apoptosis detection. EPZ5676 induced a higher percentage of apoptotic cells in the OCI-AML3 line than in the OCI-AML2 line on both day 3 and day 7 (Figure 5B). Additionally, a decrease in HOXA9 and PBX3 expression and an increase in the expression of apoptosis-related proteins and the number of apoptotic cells were detected only in KG-1 cells stably transduced with mutant NPM1 but not in those transduced with the vector or WT NPM1 after 7 days of treatment with EPZ5676 (Figure 5C-D). Reference: Theranostics. 2018 Jul 30;8(16):4359-4371. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134928/ |
| In vivo activity: | Mononuclear cells (MNCs) accumulated significantly in the liver and spleen of P. acnes-primed control mice (Figure 2A), and both the percentage and absolute number of CD4+ T cells increased dramatically in the liver and spleen (Figure 2B and C). By contrast, infiltration of MNCs and CD4+ T cells decreased significantly both in the liver and spleen of EPZ-5676–treated mice (Figure 2A–C). To detect the proliferation of CD4+ T cells, the study injected BrdU (5-bromo-2-deoxyuridine), a synthetic nucleoside that could be incorporated into newly synthesized DNA to monitor the cell proliferation, into EPZ-5676–treated or P. acnes-primed control mice. The flow cytometric analysis revealed that EPZ-5676 treatment decreased the frequencies of BrdU+ CD4+ T cells in the liver and spleen after P. acnes priming (Figure 2D), suggesting that Dot1L inhibition may regulate the proliferation of CD4+ T cells in vivo. In addition, EPZ-5676–treated mice showed decreased CD44hiCD62Llo CD4+ T cells, and increased CD62LhiCD44lo CD4+ T cells (Figure 2E), suggesting that Dot1L inhibition suppressed CD4+ T cell activation in vivo. The study also found reduced expression of chemokine receptors, such as CXCR3 and CCR7 on CD4+ T cells (Figure 3A and B) and their respective chemokines CXCL9, CXCL10, and CCL21 in the liver of EPZ-5676–treated mice (Figure 3C). These results indicated that Dot1L inhibition also suppressed the chemotaxis of pathogenic CD4+ T cells into the liver. Reference: Cell Mol Gastroenterol Hepatol. 2021 Jan 23;12(1):81-98. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8081916/ |
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 57.6 | 102.36 | |
| DMF | 30.0 | 53.31 | |
| Ethanol | 65.0 | 115.51 | |
| Ethanol:PBS (pH 7.2) (1:8) | 0.1 | 0.18 |
The following data is based on the product molecular weight 562.71 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 1 mM | 1.15 mL | 5.76 mL | 11.51 mL |
| 5 mM | 0.23 mL | 1.15 mL | 2.3 mL |
| 10 mM | 0.12 mL | 0.58 mL | 1.15 mL |
| 50 mM | 0.02 mL | 0.12 mL | 0.23 mL |
| Formulation protocol: | 1. Godfrey L, Crump NT, Thorne R, Lau IJ, Repapi E, Dimou D, Smith AL, Harman JR, Telenius JM, Oudelaar AM, Downes DJ, Vyas P, Hughes JR, Milne TA. DOT1L inhibition reveals a distinct subset of enhancers dependent on H3K79 methylation. Nat Commun. 2019 Jun 26;10(1):2803. doi: 10.1038/s41467-019-10844-3. PMID: 31243293; PMCID: PMC6594956. 2. Zhang W, Zhao C, Zhao J, Zhu Y, Weng X, Chen Q, Sun H, Mi JQ, Li J, Zhu J, Chen Z, Pandolfi PP, Chen S, Yan X, Xu J. Inactivation of PBX3 and HOXA9 by down-regulating H3K79 methylation represses NPM1-mutated leukemic cell survival. Theranostics. 2018 Jul 30;8(16):4359-4371. doi: 10.7150/thno.26900. PMID: 30214626; PMCID: PMC6134928. 3. Yang W, Yu H, Huang J, Miao X, Wang Q, Wang Y, Cheng Y, He S, Zhao F, Meng L, Wang B, Qian F, Ren X, Jin M, Gu Y, Zhang Y, Cai W. Inhibition of Dot1L Alleviates Fulminant Hepatitis Through Myeloid-Derived Suppressor Cells. Cell Mol Gastroenterol Hepatol. 2021 Jan 23;12(1):81-98. doi: 10.1016/j.jcmgh.2021.01.013. Epub ahead of print. PMID: 33497867; PMCID: PMC8081916. 4. Song Z, Wei Z, Wang Q, Zhang X, Tao X, Wu N, Liu X, Qian J. The role of DOT1L in the proliferation and prognosis of gastric cancer. Biosci Rep. 2020 Jan 31;40(1):BSR20193515. doi: 10.1042/BSR20193515. PMID: 31939604; PMCID: PMC6997103. |
| In vitro protocol: | 1. Godfrey L, Crump NT, Thorne R, Lau IJ, Repapi E, Dimou D, Smith AL, Harman JR, Telenius JM, Oudelaar AM, Downes DJ, Vyas P, Hughes JR, Milne TA. DOT1L inhibition reveals a distinct subset of enhancers dependent on H3K79 methylation. Nat Commun. 2019 Jun 26;10(1):2803. doi: 10.1038/s41467-019-10844-3. PMID: 31243293; PMCID: PMC6594956. 2. Zhang W, Zhao C, Zhao J, Zhu Y, Weng X, Chen Q, Sun H, Mi JQ, Li J, Zhu J, Chen Z, Pandolfi PP, Chen S, Yan X, Xu J. Inactivation of PBX3 and HOXA9 by down-regulating H3K79 methylation represses NPM1-mutated leukemic cell survival. Theranostics. 2018 Jul 30;8(16):4359-4371. doi: 10.7150/thno.26900. PMID: 30214626; PMCID: PMC6134928. |
| In vivo protocol: | 1. Yang W, Yu H, Huang J, Miao X, Wang Q, Wang Y, Cheng Y, He S, Zhao F, Meng L, Wang B, Qian F, Ren X, Jin M, Gu Y, Zhang Y, Cai W. Inhibition of Dot1L Alleviates Fulminant Hepatitis Through Myeloid-Derived Suppressor Cells. Cell Mol Gastroenterol Hepatol. 2021 Jan 23;12(1):81-98. doi: 10.1016/j.jcmgh.2021.01.013. Epub ahead of print. PMID: 33497867; PMCID: PMC8081916. 2. Song Z, Wei Z, Wang Q, Zhang X, Tao X, Wu N, Liu X, Qian J. The role of DOT1L in the proliferation and prognosis of gastric cancer. Biosci Rep. 2020 Jan 31;40(1):BSR20193515. doi: 10.1042/BSR20193515. PMID: 31939604; PMCID: PMC6997103. |
1: Klaus CR, Iwanowicz D, Johnston D, Campbell CA, Smith JJ, Moyer MP, Copeland RA, Olhava EJ, Scott MP, Pollock RM, Daigle SR, Raimondi A. DOT1L inhibitor EPZ-5676 displays synergistic antiproliferative activity in combination with standard of care drugs and hypomethylating agents in MLL-rearranged leukemia cells. J Pharmacol Exp Ther. 2014 Sep;350(3):646-56. doi: 10.1124/jpet.114.214577. Epub 2014 Jul 3. PubMed PMID: 24993360.
2: Basavapathruni A, Olhava EJ, Daigle SR, Therkelsen CA, Jin L, Boriack-Sjodin PA, Allain CJ, Klaus CR, Raimondi A, Scott MP, Dovletoglou A, Richon VM, Pollock RM, Copeland RA, Moyer MP, Chesworth R, Pearson PG, Waters NJ. Nonclinical pharmacokinetics and metabolism of EPZ-5676, a novel DOT1L histone methyltransferase inhibitor. Biopharm Drug Dispos. 2014 May;35(4):237-52. doi: 10.1002/bdd.1889. Epub 2014 Feb 14. PubMed PMID: 24415392.
3: Daigle SR, Olhava EJ, Therkelsen CA, Basavapathruni A, Jin L, Boriack-Sjodin PA, Allain CJ, Klaus CR, Raimondi A, Scott MP, Waters NJ, Chesworth R, Moyer MP, Copeland RA, Richon VM, Pollock RM. Potent inhibition of DOT1L as treatment of MLL-fusion leukemia. Blood. 2013 Aug 8;122(6):1017-25. doi: 10.1182/blood-2013-04-497644. Epub 2013 Jun 25. PubMed PMID: 23801631; PubMed Central PMCID: PMC3739029.
